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Epilepsies are disorders with highly phenotypic pleiotropy and genetic heterogeneity,and many problems and difficulties still exist in clinical teaching.In order to improve the quality of epilepsy teaching and train high-quality medical talents with innovative thinking,the concept of precision medicine is introduced into epilepsy teaching.Typical cases with epilepsy syndromes were collected and modified to form ideal epilepsy teaching cases.Based on the cases,specific situations and teaching plan were designed,in which the problem-based learning were applied to medical students in epilepsy teaching,in order to raise their ability of academic thinking and comprehensive analysis in the clinical practice of epilepsy.
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Objective To conduct mutation screening of SCN1A 3′ untranslated region (UTR) on Dravet syndrome (DS) patients without mutations in the SCN1A coding region and promoter region, and functional analysis of the variant from DS patients.Methods Twenty-eight DS patients without mutations in the SCN1A coding region and promoter region were screened for SCN1A 3′ UTR mutations using PCR and direct sequencing.Functional analysis of the detected mutation was done via luciferase assay, mRNA stability analysis and RNA electrophoretic mobility shift assay (RNA-EMSA).Results A novo variant (c.*20A>G) in SCN1A 3′ UTR was found in one DS patient.The variant (c.*20A>G) reduced the luciferase gene xpression by 30% through increasing the affinity of pluripotent embryonal carcinoma cell line NT2/cytoplasmic protein binding and reducing luciferase gene mRNA stability (t=8.5,P<0.01).Conclusions A functional variant was detected from one patient with DS.This variant negatively regulated the gene expression by increasing the affinity of pluripotent embryonal carcinoma cell line NT2/cytoplasmic protein binding and reducing mRNA stability.
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Objective To investigate the potential pathogenesis of Focal cortical dysplasia (FCD), we performed cDNA microarray analysis to obtain gene expression profile of FCD. Methods Three FCD samples and three normal controls were enrolled. Total RNA of the brain tissues were extracted. The difference gene expressions between FCD group and control group was detected using Affymetrix gene chip. The up and down-regulated genes were confirmed by Real-time PCR. Further, the related signal pathways involved in the pathogenic mechanisms of FCD were predicted by bioinformatics. Result In FCD, two up-regulated genes C21orF2 and AU152162 and 5 down-regulated genes ENPP2, ANLN, IP6K3, UGT8, and AZGP were found. Compared the FCD samples with the normal controls , there were significantly different in all down-regulated genes (P 0.05). Using bioinformatics analysis, the ENPP2 , UGT8 , and AZGP1 protein which located in the cell membrane or secreted into the extracellular matrix may be involved in the formation of the myelin sheath and the development of the nervous system by the lipid metabolism and LPA signaling pathway. Conclusion ENPP2, UGT8 and AZGP1 may be involved in pathogenesis of FCD through the process of myelin sheath formation and LPA signal pathway , which warrants further study to know their roles in the pathogenesis of FCD.
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Objective To analyze the causes of misdiagnosis and mistreatment of Dravet syndrome. Methods Patients with Dravet syndrome diagnosed according to clinical features and SCN1A gene mutation detection were recruited within recent 3 years. The patients were grouped into correct diagnosis-treatment group and misdiagnosis-mistreatment group according to whether the patients had ever been misdiagnosed and mistreated by sodium channel blockers. The clinical features were compared between two groups. Results Thirty-five cases with Dravet syndrome were collected and the rate of misdiagnosis reached 40%, Nine cases were misdiagnosed as symptomatic focal epilepsy, 4 as Lennox-Gastaut syndrome and 1 as Doose syndrome. The average age of onset in misdiagnosis-mistreatment group was (5.50 ± 3.56) months,and the age of confirmed diagnosis was (83.57 ± 105.62) months. The percentage of abnormal EEG, onset seizure with partial seizure, the seizure frequency within the first year from onset, onset with afebrile seizure, patients with status epilepticus or cluster seizures was higher in misdiagnosis-mistreatment group but it showed no significant statistical significance when compared with that of correct diagnosis-treatment group. The percentage of patients with mental retardation and focal neurological signs was significantly higher in misdiagnosis-mistreatment group (P=0.005 and 0.002, respectively). Conclusions Dravet syndrome is frequently misdiagnosed as symptomatic focal epilepsy. The appearance of focal neurological signs and mental retardation before confirmed diagnosis are important factors for misdiagnosis. Gene mutation screening will be helpful for differential diagnosis of Dravet syndrome.
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Objective To study the clinical characteristics of autism in febrile seizures plus (FS+) and the relationship between autism and SCN1A mutation. Methods Clinical data of 103 patients with FS+ treated in epilepsy centre of the Second Affiliated Hospital of Guangzhou Medical University were collected and analyzed. According to the international criteria, generalized epilepsy with febrile seizures plus (GEFS+), partial seizures with febrile seizures plus (PEFS+), Dravet syndrome (DS) and autism were diagnosed. Genomic DNA was obtained from blood samples. SCN1A were PCR amplified and mutations were detected by DHPLC and sequencing. Result Mental retardation was found in 53.8%of patients with GEFS+, 69.2%of patients with PEFS+, and all patients with DS, respectively. One in GEFS+, one in PEFS+and nine in DS patients were accompanied with autism (P<0.01). Among FS+patients with autism, one SCN1A mutation was found in PEFS+patients, while six SCN1A mutations were found in DS patients. Conclusions Majority of GEFS+and PEFS+patients showed mental retardation, while all the DS patients were accompanied with retardation. The occurrence of autism with DS is higher than GEFS+and PEFS+. No definite relationship between autism and SCN1A mutation was indicated.
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Objective To screen and analyze nucleotide variants in 5'-untranslated region(5'-UTR)in voltage-gated sodium channel α1-subunit gene(SCN1A)in patients with Dravet syndrome and to evaluate the association of the variants with disease.Methods Peripheral blood of 24 patients with Dravet syndrome and 100 unrelated normal persons were collected and genomic DNA was extracted.PCR-sequencing of SCN1 A 5'-UTR in these DNA was performed.To evaluate the possibility of mutation inducing disease,bioinformatics analysis was applied to analyze the conservation of the sequences around the mutation site and predict the potential transcription elements.Results The nucleotide variant of 166.642.520G→A in exon 2 was identified in two patients,but not in normal controls.The mutation was a de novo mutation in a patient with early-onset.In the second proband,the mutation was also carried by his clinically asymptomatic mother.The nucleotide site 166.642.520 was moderately conserved in mammals(62.5%).The average nucleotide identity rate between human and other mammals species in the region adjacent to 166.642.520 was 88.5%.Two potential transcription regulatory elements were predicted on the sequence with the mutation of 166.642.520G>A,and only one on the sequence with wild-type.Conclusions The mutation 166.642.520G>A may be associated with Dravet syndrome and further studied should be performed to verify it and demonstrate its pathogenic mechanisms.
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@#Objective To investigate the neuroprotective effect of extraction of gastrodia elata on status epilepticus induced by Li-pilocarpine in rats. Methods 144 SD rats were randomly divided into 4 groups: saline control (5 ml/kg twice daily), small dose gastrodia group (5 ml/kg twice daily), large dose gastrodia group (10 ml/kg twice daily) and lamotrigine group (20 mg/kg twice daily). The status epilepticus was induced 3 d after treatment. The duration of status epilepticus, neuron loss, the rate of spontaneous seizure and the score of mossy fiber sprouting were analyzed. Results Large dose gastrodia or lamotrigine administration can shorten the duration of status epilepticus and decrease the neuron loss compared with the small dose gastrodia and control administration. But all treatments cannot prevent mossy fiber sprouting and spontaneous seizures. Conclusion 20 ml/kg gastrodia or 40 mg/kg lamotrigine administration can shorten the duration of status epilepticus induced by Li-pilocarpine, and decrease the neuron loss of CA1, CA3 of hippocampus and hilus of dentate gyrus, but cannot prevent epileptogenesis after brain insult.
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Objective To study the sodium channel α1-subunit (SCN1A) gene in a pair of monozygotic twins with borderland severe myoclonic epilepsy in infancy (SMEB) and its characteristic of clinical manifestations. Methods The clinical features of 2 monozygotic twins were summarized. All 26 exons of SCNIA genes were screened with denaturing high performance liquid chromatography (DHPLC), and direct sequence analysis was performed on those with abnormal elution peak. Results The proband and her sister showed typical clinical features of SMEB. The same heterozygous mutations on exon 26 which caused the related amino acid change were found among them (c. 5348C > T, A1783E). Conclusion Monozygotic twins with similar clinical phenotype of SMEB have same SCN1A gene mutation.
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Objective To explore the inheritance characteristics of SCN1A gene in familial severe myoclome epilepsy in infancy.Methods The clinical information and blood of the patients and their relatives who had febrile seizure(FS)or epilepsy history were collected.Blood genome DNA were extracted.All exons of SeN1A gene were PCR amplified and screened with denaturing high Performance liquid chromatography(DHPLC)technology,and sequence analysis was performed.Results Fourteen SME patients had FS or epilepsy family history.Five were found positive history in first class relatives and 2 of them had inherited mutations of SCN1A(C.4284+2T>C and e.1216G>T):Other9 were found positive history in second class relatives and 2 of them had de novo mutations of SCN1A.Condusions SCN1A is the pathogenic gene for SME.The same muatation of SCN1A gene can be related to different clinical phenotypes.SME patients whose first class relatives with FS or epilepsy history should be taken as the focus of SCN1A inherited mutation screening.
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Objective To study the SCN1A gene in a family with partial epilepsy with febrile seizures plus ( PEFS+ ) and its characteristics of inheritance. Methods The clinical features of the 2 patients and their father were summarized. All 26 exons of SCN1A gene were screened with denaturing high performance liquid chromatography (DHPLC), and direct sequence analysis was pedormed on those with abnormal elution peak. Pyrosequencing was subsequently performed in those without abnormality in direct sequence analysis. Results The proband and his sister had the phenotype of PEFS+ . The same heterozygous mutations (AS768G) on exon 26 which caused the related amino acid change (Q1923R) were found among them. Their father had frequent febrile seizures (FS) in childhood, and seizures stopped spontaneously. No abnormality was found in direct sequence but mosaic mutation in the same site was discovered with pyrosequencing (mutation quantity was 25% ). Conclusions The mutatin of SCN1A could cause partial epilepsy. PEFS+ could be inherited, the relatives carrying the affected gene may have mild clinical symptoms, possibly resulting from the low concentration of the mutated gene due to mosaic mutation.
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@#Objective To explore the role of Kv channel interacting protein 1(KChIP1)in the process of epileptic seizure and the relationship between KChIP1 and gamma-aminobutyric acid-(GABA)ergic neurons.Methods Normal female Sprague-Dawley rats were treated with pentylenetetrazole to make acute pentylenetetrazole models of epilepsy.Laser Scanning Confocal Microscope(LSCM)combined with double-labeled immunohistochemical technique was applied to observe the expression of the KChIP1 and the GABAergic neurons in the hippocampus of rats.Results The number of KChIP1-postive neurons in the hippocampus was significantly increased in the acute pentylenetetrazole model rats(P<0.05).There was no significant difference in the number of double-labled neurons(P>0.05),nor of the GABA-postive neurons between the model rats and the controls.The ratio of double-labeled neurons/total positive neurons was 63.9% in the hippocampus.Conclusion The KChIP1 might be involved in epileptogenesis of pentylenetetrazole induced seizure.The KChIP1 was associated with GABAergic neurons,whereas it may be functionally different from GABA.
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In order to improve the quality of epilepsy teaching,the appropriate version of the international classification of epilepsy and epileptic syndrome were selected to teach in the different level students by the way of PBL and clinical case analysis.The clinical thoughts and enthusiasm were improved.The classification of epilepsy could be grasped and easily used in their clinical work.
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Generalized epilepsy with febrile seizures plus is a novel and common generalized epilepsy syndromes, with significant genetic and phenotypic heterogeneity. Research on genetic position and mutant of generalized epilepsy with febrile seizures plus has been a hot spot, which has importance in clarifying epilepsy syndrome as a kind of channelopathy.
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Objectives To evaluate the valproate plasma l ev els,the clinical efficacy and adverse effects in patients with epilepsy treated with the conventional preparations and the sustained-release preparations of sodium valproate (VPA-Na?SR).Methods 33 patients received oral conventional formul ation of sodium valproate for over six months and a similar dosage of VPA-Na?SR for 4 weeks.After 12 or 24 hours,the valproate plasma concentr ations of the two formulations were measured respectively before taking drugs in the early mor ning.The valproate plasma concentrations and the clinical efficacy of the VPA-N a?SR were assessed by comparing with that of conventional valproate.The adverse effects were recorded.Results The average valproate plasma trough concentration was s ignificantly higher in patients receiving VPA-Na?SR than that of those receiving conventional valproate.Seizure free in patients was achi eved by 76%(n=25) with VPA-Na?SR and by 45%(n=15) with conventional valproate resp ectively.There was statistical difference between the two formulations.The seizu re frequency was significantly reduced in 5 patients treated with VPA-Na?SR.A dverse ef fects were observed in 2 patients with conventional valproate,5 patients with V PA-Na?SR whose valproate plasma levels were higher than that of conventional p reparations.Adverse effects were related to increased valproate plasm a levels and individual drugtolerance. Conclusions The advantage of VPA-Na?SR is that serum valproa te con centrations may increase smoothly and minimize fluctuation in serum dr ug concentrations during a dosing interval. It is a more effective and more convenient antiepileptic agent.
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Objective To investigate the efficacy and safety of Topiramate as adjunctive therapy in patients with refractory epilepsy.Methods 115 patients enrolled in this open-label,self-controlled prospective study which were designed to adopt slower dosage escalation schedule with lower initial,until reached target dosages(up to 8 weeks) or the most tolerable dosages of Topiramate,and maintained them 1~3 years treatment.The long-term efficacy,safety of Topiramate and their relationship with dosage were evaluated.Results The median percentages reduction were 56.0%,75.8%,76.1%,77.3% and 78.1% after 8 weeks,6 months,1 year,2 years and 3 years Topiramate treatment respectively for adults,and 32.1%,66.7%,68.9%,70.1% and 70.8% respectively for children.The overall efficacy for partial seizures(70.6%) was higher than that for generalized seizures(37.5%)((P
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Objective To observe the effect of synthetic corticotropin with vitamin B 6 on intractable epilepsy of children.Methods 20 patients (3 months to 4 years old) among which 9 children with infantile spasms and 11 children with intractable epilepsy were injected with vitamin B 6 (50～100 mg/d) for 10 days,then treated with cortrosyn 0 015~0 025 mg/Kg?d by the interval mothod of lengthening dosage,total course of treatment was about 2 months,seizure,EEG and some blood biochemical tests were observed in all patients before and after the course of treatment.Results The seizure of 14 patient children stopped after the treatment,seizure frequency was decreased in 5 patients (from 15 to 40 times/week decreasing to 3 to 14 times/week respectively),1 patient with intractable epilepsy was stopped treating because of side effect.The general clinical condition and EEG were also improved.The blood biochemical tests about liver and kidney were normal during treatment,hypokaliemia,hypocalcemia and slight dropsy were appeared in some patients.Conclusion Cortrosyn with vitamin B 6 is effective for infantile spasms and intractable epilepsy in children of low age.
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Objective To investigate the role of proton magnetic resonance spectroscopy (~1H-MRS) in accurate lateralization of temporal lobe epilepsy (TLE) and the factors which influenc ~1H-MRS in epileptogenic zones.Methods ~1H MRS and MRI were performed in 40 patients with TLE and 20 healthy volunteers by 1.5T MR/MRS system. The data of spectra of N-acetylaspartate (NAA), creatine+phosphocreatine (Cr) and choline-containing compounds (Cho) obtained from the medial regions of the temporal lobes were analyzed. Lateralization of TLE with ~1H-MRS was defined by the ratio of NAA/(Cho+Cr).Results The ratios of NAA/(Cho+Cr) both in epileptogenic zones (0.45?0.12) and contralateral regions (0.51?0.10) were lower than that in control group (0.58?0.09)( P0.05). There was significant difference in bilateral abnormal EEG between bilateral abnormal NAA/(Cho+Cr) and unilateral abnormal NAA/(Cho+Cr)( P
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10 mm in diameter and tended to be correlated with the main discharges in EEG in two cases. 11 of 20 cases (61.11%) were effective with AEDs. After more than one year following up, 3 of 10 patients became worse.Conclusion Seizures in TS usually begin in young children, and partial seizures is the commonest seizure type in this study. There is a shift in seizure type during the clinical course. The results also suggests that there is a certain correlation between the size of cortical tubers and the epiletogenic focus. Treatment with AEDs is effective. However, the effect appears decreasing with increasing age.
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Objective To explore the effect of astragalus polysaccharides (APS) and soy isoflavones (SI) on glucose metabolism in diabetic rats. Methods Healthy 60 rats were equally and randomly divided into 5 groups, normal forage group (NF), high blood sugar model group(HBS), APS high dose group intervention group (APS-H), APS low dose group (APS-L), and SI intervention group (SI). Diabetic model was inflicted by intraperitoneal injection of streptozotocin (STZ). Ordinary feed and APS high dose,low dose APS and SI interfere were respectively given for 8 weeks. Blood glucose,glycated hemoglobin,glucose and lipid levels were determined in every group. Results After APS high dose,low dose and SI were given to feed diabetic rats for 8 weeks, their blood glucose, glycated hemoglobin, glucose, TC, TG, LDL, VLDL and high blood glucose was significantly lower than the HBS group (P